Rapid online proteolytic mapping of PEGylated rhGH for identity confirmation, quantitation of methionine oxidation and quantitation of UnPEGylated N-terminus using HPLC with UV detection.

Proteolytic mapping is a widely used tool in the BioPharmaceutical Industry for the analysis of post-translation modifications as well as confirmation of protein identity by comparison to a well-characterized reference standard. This manuscript presents an integrated chromatographic approach which provides the ability to rapidly digest and analyze a PEGylated rhGH for methionine oxidation, identity confirmation and free (unPEGylated) N-terminal peptide by RP-HPLC using UV detection at 280 nm. This approach utilizes an online procedure in which the digestion step is integrated to the RP-HPLC analysis via an external column switching valve.

High binding capacity surface grafted monolithic columns for cation exchange chromatography of proteins and peptides.

Poly(glycidyl methacrylate-co-ethylene methacrylate) monoliths have been prepared in 100 microm i.d. capillaries and their epoxy groups hydrolyzed to obtain poly(2,3-dihydroxypropyl methacrylate-co-ethylene methacrylate) matrix.